Materials and methods for the delivery of biomolecules to cells of an organ

Abstract

The present invention is directed towards materials and methods of delivering biomolecules to cells that define an organ, and the organ being in situ, and electroporating cells defining the organ by delivering to the cells an amount of energy effective to electroporate the cells. The present invention is also directed towards materials and methods of expressing heterologous polypeptides in organs of a subject and electroporating cells defining the organ by delivering to the cells an amount of energy effective to electroporate the cells.

1 . A method of delivering a biomolecule to a cell that defines an organ, the organ being in situ, comprising:
 delivering the biomolecule to the organ, and   providing an amount of energy to the organ effective to electroporate the cells that define the organ.   
     
     
         2 . The method of  claim 1 , wherein the delivering step further comprises:
 providing a pharmaceutical formulation comprising the biomolecule for delivery to a subject having the organ.   
     
     
         3 . The method of  claim 2 , wherein the delivering step further comprises:
 preparing a delivery apparatus with the pharmaceutical formulation.   
     
     
         4 . The method of  claim 1 , wherein the providing an amount of energy step further comprises:
 situating at least one pair of electrodes around the organ,   generating the electroporating amount of energy, and   transmitting the electroporating amount of energy to the pair of electrodes.   
     
     
         5 . The method of  claim 1 , wherein the biomolecule is a polynucleic acid, polypeptide, or pharmaceutical compound. 
     
     
         6 . The method of  claim 5  wherein the polynucleic acid is a siRNA, linear DNA, or DNA expression construct. 
     
     
         7 . The method of  claim 5  wherein the polypeptide is a native protein, heterologous polypeptide, or protein fragment. 
     
     
         8 . The method of  claim 5 , wherein the polynucleic acid is a DNA expression construct comprising an encoding sequence. 
     
     
         9 . The method of  claim 8 , wherein the encoding sequence encodes GHRH, IGF-1, caspase, or cytokine. 
     
     
         10 . The method of  claim 8  wherein the encoding sequence encodes GHRH. 
     
     
         11 . The method of  claim 8 , wherein the DNA expression construct comprises a tissue specific promoter operably linked to the encoding sequence. 
     
     
         12 . The method of  claim 11  wherein the tissue specific promoter is SPc5-12, SPc1-26, or SPc6-39. 
     
     
         13 . The method of  claim 1 , comprising transdermal, transthoracic, or direct delivery of the biomolecule to the organ. 
     
     
         14 . The method of  claim 1 , further comprising:
 imaging the delivery of the biomolecule to the organ.   
     
     
         15 . The method of  claim 14  wherein the imaging is ultrasound imaging. 
     
     
         16 . A method of expressing a heterologous polypeptide in an organ of a subject, comprising:
 injecting a needle transdermally, transthoracically, or directly to the organ for delivery of an encoding nucleic acid molecule that encodes the heterologous polypeptide, and   electroporating cells defining the organ by delivering to the cells an amount of energy effective to electroporate the cells.   
     
     
         17 . The method of  claim 16 , further comprising:
 providing a pharmaceutical formulation comprising the encoding nucleic acid for delivery to the organ, and   preparing a delivery apparatus with the pharmaceutical formulation, said delivery apparatus being in fluid communication with said needle.   
     
     
         18 . The method of  claim 16 , wherein the electroporating step further comprises:
 situating at least one pair of electrodes around the organ,   generating the electroporating amount of energy, and   transmitting the electroporating amount of energy to the electrodes.   
     
     
         19 . The method of  claim 16 , wherein the encoding nucleic acid encodes GHRH, IGF-1, caspase, or cytokine. 
     
     
         20 . The method of  claim 16 , wherein the encoding nucleic acid
 comprises a tissue specific promoter operably linked to the encoding sequence.   
     
     
         21 . The method of  claim 20 , wherein the tissue specific promoter is SPc5-12, SPc1-26, or SPc6-39. 
     
     
         22 . The method of  claim 16 , further comprising:
 imaging the delivery of the biomolecule to the organ.   
     
     
         23 . The method of  claim 22  wherein the imaging is ultrasound imaging. 
     
     
         24 . A system adapted for the delivery of an encoding nucleic acid molecule to an organ of a mammal for the expression of a polypeptide from cells defining the organ, comprising:
 a needle for delivery of the encoding nucleic acid molecule to the organ, and   an electorporation device capable of delivering pulses of energy at an intensity effective for electroporation of the cells of the organ.   
     
     
         25 - 28 . (canceled)